High cell-density fermentation of shark hepatical stimulator analogue in Escherichia coli / 生物工程学报
Chinese Journal of Biotechnology
; (12): 1371-1378, 2009.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-296914
Responsible library:
WPRO
ABSTRACT
The potential effects of recombinant shark hepatical stimulator analogue (r-sHSA) in liver disease have been revealed in our previous studies. In order to further evaluate its clinic application, we carried out high cell-density fermentation in 5 L fermentor to get enough products. Based on the trials in shaking flask, we optimized the parameters for 5 L fermentor, including medium composition, medium supplement, inducer concentration and induction time, etc. In detail, the improved LB medium (0.97% glycerol, 0.91% yeast extract, 0.72% tryptone, 0.782% KH2PO4, 0.267% K2HPO4.3H2O, 0.062% MgSO4.7H2O, 0.5% NaCl, pH 7.0) is chosen to cultivate the engineering bacteria with the constant fermentation condition (pH 7.0, and the dissolved oxygen concentration is about 25%-30%). When bacterial culture reaches exponential phase, the modified feeding medium (620 g/L glycerol, 94.8 g/L tryptone, 3.3 mL/L trace elements, and 7.5 g/L MgSO4.7H2O) is then supplied through the method of exponential fed-batch mode. After the optical density (OD600) of engineering bacterial culture reaches to 23, the ultimately concentration of 0.5 mmol/L IPTG is added to induce the expression of r-sHSA for 6 h. Results show that the amount of r-sHSA production is (2.662 +/- 0.041) g/L, which is about 13.7 folds of the one optimized before.
Full text:
Available
Health context:
Neglected Diseases
Health problem:
Neglected Diseases
/
Zoonoses
Database:
WPRIM (Western Pacific)
Main subject:
Peptides
/
Sharks
/
Recombinant Proteins
/
Chemistry
/
Cloning, Molecular
/
Escherichia coli
/
Fermentation
/
Genetics
/
Liver
/
Metabolism
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2009
Document type:
Article